Maura Ferri

Chitosan treatment induces changes of protein expression profile and stilbene distribution in Vitis vinifera cell suspensions

Polyphenols, including stilbenes and flavonoids, are an essential part of human diet and constitute one of the most abundant and ubiquitous groups of plant secondary metabolites, and their level is inducible by stress, fungal attack or biotic and abiotic elicitors. Proteomic analysis of Vitis vinifera (L.) cultivar (cv.) Barbera grape cell suspensions, showed that the amount of 73 proteins consistently changed in 50 μg/mL chitosan‐treated samples compared with controls, or between the two controls, of which 56 were identified by MS analyses. In particular, de‐novo synthesis and/or accumulation of stilbene synthase proteins were promoted by chitosan which also stimulated trans‐resveratrol endogenous accumulation and decreased its release into the culture medium. No influence was shown on cis‐resveratrol. There was no effect on the accumulation of total resveratrol mono‐glucosides (trans‐ and cis‐piceid and trans‐ and cis‐resveratroloside). Throughout the observation period the upregulation of phenylalanine ammonia lyase, chalcone synthase, chalcone‐flavanone isomerase (CHI) transcript expression levels well correlated with CHI protein amount and with the accumulation of anthocyanins. Chitosan treatment strongly increased the expression of eleven proteins of the pathogenesis related protein‐10 family, as well as their mRNA levels.

Resveratrol production in Vitis viniferacell suspensions treated with several elicitors

Abstract We investigated the effect of several elicitors on the production of resveratrol in Vitis vinifera cv. Barbera cell suspension cultures. Salicylic acid, Na-orthovanadate, jasmonates, chitosan and the monomers D-glucosamine and N-acetyl-D-glucosamine, ampicillin and rifampicin differently elicitated the accumulation of the trans and cis resveratrol isomers, both in the cells and in the culture media. Proteomic analyses also showed that methyl-jasmonate and chitosan stimulated the ex-novo synthesis of the stilbene synthase protein.

Recovery of polyphenols from red grape pomace and assessment of their antioxidant and anti-cholesterol activities.

The present work aimed at the recovery and characterization of polyphenolic compounds extracted from red grape pomace (Vitis vinifera L.), a winemaking by-product. Polyphenolic compounds of wet (WP) and dried (DP) red pomace were recovered by enzymatic digestions and ethanol-based extractions. Fungamyl and Celluclast enzymes were found to be the most effective in enhancing polyphenol release from WP. WP samples showed the highest capacity of releasing polyphenols with 2h control 24°C and 2h 1% Celluclast resulting as the best treatments. A significantly lower amount of polyphenols was recovered from DP most probably as a consequence of the pomace drying. The best extracts contained high amounts of total polyphenols, flavonoids, tannins and anthocyanins and exerted antioxidant and cholesterol-lowering activities. The results support the possibility of exploiting the extracts coming from grape processing by-products as ingredients for functional and innovative products in the nutraceutical, pharmaceutical or cosmetic fields.

Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity

AbstractThe availability of smartphones with high-performance digital image sensors and processing power has completely reshaped the landscape of point-of-need analysis. Thanks to the high maturity level of reporter gene technology and the availability of several bioluminescent proteins with improved features, we were able to develop a bioluminescence smartphone-based biosensing platform exploiting the highly sensitive NanoLuc luciferase as reporter. A 3D-printed smartphone-integrated cell biosensor based on genetically engineered Hek293T cells was developed. Quantitative assessment of (anti)-inflammatory activity and toxicity of liquid samples was performed with a simple and rapid add-and-measure procedure. White grape pomace extracts, known to contain several bioactive compounds, were analyzed, confirming the suitability of the smartphone biosensing platform for analysis of untreated complex biological matrices. Such approach could meet the needs of small medium enterprises lacking fully equipped laboratories for first-level safety tests and rapid screening of new bioactive products. Graphical abstractSmartphone-based bioluminescence cell biosensor

Ethnobotanical survey of wild food plants traditionally collected and consumed in the Middle Agri Valley (Basilicata region, southern Italy)

BackgroundThis research was carried out in a scarcely populated area of the Middle Agri Valley (Basilicata region, southern Italy). The aim of the study was to record local knowledge on the traditional uses of wild food plants, as well as to collect information regarding the practices (gathering, processing and cooking) and the medicinal uses related to these plants.MethodsFifty-eight people still possessing traditional local knowledge (TLK), 74% women and 26% men, were interviewed between May–August 2012 and January 2013, using open and semi-structured ethnobotanical interviews. For each described plant species, the botanical family, the Italian common and folk names, the plant parts used, the culinary preparation and, when present, the medicinal use, were recorded and the relative frequency of citation index (RFC) was determined.ResultsThe 52 plant species mentioned by the respondents belong to 23 botanical families, with Asteraceae (12 plants) and Rosaceae (7 plants) being most frequently cited. The species with the highest RFC index is Cichorium intybus L. (0.95), followed by Sonchus spp. (S. oleraceus L., S. asper L. and S. arvensis L.) (0.76). The plant parts preferably used are leaves (22 plants), fruits (12) and stems (7). Only six wild plants were indicated as having both food use and therapeutic effect.ConclusionsThe survey conducted on the traditional use of wild food plants in the Middle Agri Valley revealed that this cultural heritage is only partially retained by the population. Over the last few decades, this knowledge has been in fact quickly disappearing along with the people and, even in the rural context of the study area, is less and less handed down to younger generations. Nevertheless, data also revealed that the use of wild plants is recently being revaluated in a way closely related to local habits and traditions.

Capsicum spp in vitro liquid cell suspensions: A useful system for the production of capsaicinoids and polyphenols

Abstract Capsicum are among the most extensively cultivated and consumed plant species in the world, because of their unique pungency, aroma and colour. The typical burning sensation caused by chili peppers is due to the occurrence of a group of alkaloids named capsaicinoids. In the present study, the production of solid callus and cell suspensions from hypocotyl explants of three different chili pepper cultivars (Capsicum annuum L. cv. Mazzolino, Capsicum chinense Jacq. cv. Naga Morich and Pimenta de Neyde), was optimised. In addition, C. chinense cv. Naga Morich cell suspensions were supplemented with biotic elicitors (methyl-jasmonate and chitosan) and with precursors and intermediates of capsaicin biosynthesis (vanillin, phenylalanine and valine), and both cells and media were analysed for capsaicinoid, polyphenol, flavonoid contents and for antioxidant activity. This is the first report regarding capsaicinoid elicitation with pure chitosan and with a combination of precursors of both phenylpropanoid and valine pathways. Overall, the highest capsaicinoid levels were detected in cell extracts from cultures treated with 10 μM methyl-jasmonate and with a combination of phenylalanine and valine amino acids (100 μM each). The present results confirm the possibility of using hypocotyl chili pepper cell suspensions to produce high amounts of health beneficial metabolites.

White grape pomace extracts, obtained by a sequential enzymatic plus ethanol-based extraction, exert antioxidant, anti-tyrosinase and anti-inflammatory activities

The present work aimed at optimizing a two-step enzymatic plus solvent-based process for the recovery of bioactive compounds from white grape (Vitis vinifera L., mix of Trebbiano and Verdicchio cultivars) pomace, the winemaking primary by-product. Phenolic compounds solubilised by water enzyme-assisted and ethanol-based extractions of wet (WP) and dried (DP) pomace were characterised for composition and tested for antioxidant, anti-tyrosinase and anti-inflammatory bioactivities. Ethanol treatment led to higher phenol yields than water extraction, while DP samples showed the highest capacity of releasing polyphenols, most probably as a positive consequence of the pomace drying process. Different compositions and bioactivities were observed between water and ethanol extracts and among different treatments and for the first time the anti-tyrosinase activity of V. vinifera pomace extracts, was here reported. Enzymatic treatments did not significantly improve the total amount of solubilised compounds; Celluclast in DP led to the recovery of extracts enriched in specific compounds, when compared to control. The best extracts (enzymatic plus ethanol treatment total levels) were obtained from DP showing significantly higher amounts of polyphenols, flavonoids, flavanols and tannins and exerted higher antioxidant and anti-tyrosinase activities than WP total extracts. Conversely, anti-inflammatory capacity was only detected in water (with and without enzyme) extracts, with WP samples showing on average a higher activity than DP. The present findings demonstrate that white grape pomace constitute a sustainable source for the extraction of phytochemicals that might be exploited as functional ingredients in the food, nutraceutical, pharmaceutical or cosmetic industries.

Bioprospecting on invasive plant species to prevent seed dispersal

AbsractThe most anthropized regions of the world are characterized by an impressive abundance of invasive plants, which alter local biodiversity and ecosystem services. An alternative strategy to manage these species could be based on the exploitation of their fruits in a framework of bioprospecting to obtain high-added value compounds or phytocomplexes that are useful for humans. Here we tested this hypothesis on three invasive plants (Lonicera japonica Thunb., Phytolacca americana L., and Prunus serotina Ehrh.) in the Po plain (northern Italy) which bear fruits that are highly consumed by frugivorous birds and therefore dispersed over large distances. Our biochemical analyses revealed that unripe fruit shows high antioxidant properties due to the presence of several classes of polyphenols, which have a high benchmark value on the market. Fruit collection for phytochemical extraction could really prevent seed dispersal mediated by frugivorous animals and produce economic gains to support local management actions.

Peptide Fractions Obtained from Rice By-Products by Means of an Environment-Friendly Process Show In Vitro Health-Related Bioactivities.

Recently, the isolation of new health-related bioactive molecules derived from agro-food industrial by-products by means of environment-friendly extraction processes has become of particular interest. In the present study, a protein by-product from the rice starch industry was hydrolysed with five commercial proteolytic enzymes, avoiding the use of solvents or chemicals. The digestion processes were optimised, and the digestates were separated in fractions with four different molecular weight ranges by using a cross-flow membrane filtration technique. Total hydrolysates and fractions were tested in vitro for a wide range of biological activities. For the first time rice-derived peptides were assayed for anti-tyrosinase, anti-inflammatory, cytotoxicity and irritation capacities. Antioxidant and anti-hypertensive activities were also evaluated. Protamex, Alcalase and Neutrase treatments produced peptide fractions with valuable bioactivities without resulting cytotoxic or irritant. Highest levels of bioactivity were detected in Protamex-derived samples, followed by samples treated with Alcalase. Based on the present results, a future direct exploitation of isolated peptide fractions in the nutraceutical, functional food and cosmetic industrial fields may be foreseen.

Publisher Correction: Bioprospecting on invasive plant species to prevent seed dispersal

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